Biotechnology Journal
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2021
·Vol. 17
·期 1
·2100335
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IF 4.677
Shiyu He,Yanzhi Huang,Yingwei Ma,Haoyan Yu,Bo Pang,Xingxing Liu,Caihong Yin,Xiaomu Wang,Yuan Wei,Yuling Tian,Chao Zhao,Kun Xu,Juan Wang,Chunping Lv,Xiuling Song,Minghua Jin
Foodborne pathogen contamination is a major safety issue for many foods and is causing concern worldwide. In this study, a detection system based on magnetic separation, multiplex PCR (MPCR) and capillary electrophoresis (CE) technologies was developed for the simultaneous detection of four foodborne pathogens. Magnetic separation technology is used to rapidly capture pathogenic bacteria in food samples, and then a combination of MPCR and CE can be used to greatly increase detection sensitivity. The detection limit for bacterial DNA reached 10 −5 –10 −7 ng μL −1 and in the analysis of mocked food samples, the assay showed good sensitivity for bacterial detection ranging from 10 1 to 10 5 CFU mL −1 with excellent specificity. Compared to similar detection methodologies, this technique avoids the need for time-consuming enrichment cultures, is more sensitive, and can be used to assay simultaneously four foodborne pathogens. Graphical and Lay Summary Food-borne bacteria contamination is one of the main problems of food safety, which seriously affects the economic development of many countries. In this study, the author achieved rapid separation and enrichment of multiple pathogens in a complex sample matrix through magnetic nano carbon dots, and constructed a rapid detection system for four foodborne pathogens by combining multiplex PCR and capillary electrophoresis technologies. This work shows great potential and application value in food safety monitoring.
研究领域:
食品科学
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微生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
Applied Sciences-Basel
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2022
·Vol. 12
·期 13
·6415
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IF 2.838
Fangbin Xiao,Xuekun Bai,Keyu Wang,Yifan Sun,Hengyi Xu
Featured ApplicationRapid magnetic enrichment strategy can significantly improve the sensitivity of multiplex PCR analysis and shorten the pre-concentration time. The simultaneous and sensitive detection of pathogenicListeriaspp. was achieved.AbstractListeria monocytogenesandListeria ivanoviiare important pathogenicListeriaspp. that cause infections in humans and animals. Establishing a rapid and sensitive method for the simultaneous screening of pathogenicListeriaspp. is of great significance for ensuring food safety. Multiplex polymerase chain reaction (mPCR) has been extensively reported to simultaneously detect several pathogens in food with high sensitivity, but a time-consuming pre-enrichment process is necessary. In this study, we report the usage of surface-modified polyethyleneimine-coated positively charged magnetic nanoparticles (PEI-MNPs) for rapid enrichment of pathogenicListeriaspp. through electrostatic interactions. The enrichment process takes only 10 min with high capture efficiency (more than 70%) at a wide pH range and ionic strength. Combined with mPCR analysis, the PEI-MNPs-mPCR strategy can simultaneously, rapidly, and sensitively detect pathogenicListeriaspp. without a time-consuming pre-concentration process. Under the optimal conditions, the detection limits ofL. monocytogenesandL. ivanoviiin lettuce were both as low as 101CFU/mL, which was a hundred times lower than that without magnetic enrichment. In conclusion, the magnetic enrichment strategy based on charge interaction combined with mPCR analysis has great application potential in shortening the pre-concentration time of foodborne pathogens and improving the detection sensitivity.Keywords:magnetic enrichment;multiplex PCR;electrostatic interaction;bacteria detection;method sensitivity;food safety
研究领域:
食品科学
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微生物学
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分子生物学
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产品关联
DOI
高影响因子
Food Analytical Methods
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2021
·Vol. 14
·期 9
·1964-1976
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IF 3.366
Chen Guanhua,Yang Guotai,Wang Yutong,Deng Mei,Wang Zhengzheng,Aguilar Zoraida P.,Xu Hengyi
Sensitive, simple, and rapid pathogen separation and detection methods are essential for the early diagnosis and immediate medical treatment of bacterial infections. In this study, a novel strategy based on vancomycin (Van)-modified ovalbumin (OVA) mediated magnetic nanoprobes (MBs-OVA-Van) combined with multiplex polymerase chain reaction (mPCR) was developed for efficient isolation and detection of Staphylococcus aureus ( S. aureus ) and Bacillus cereus ( B. cereus ). Van conjugated to ovalbumin coated magnetic microcarrier via strong covalent interaction. In addition, Van binds through hydrogen bonds with d -alanyl- d -alanine moieties on the surface of gram-positive bacteria cell wall. Under optimum conditions, the MBs-OVA-Van showed that the capture efficiency (CE) for S. aureus and B. cereus in PBS and juice samples were around 91%, 92% and 89%, 85%, respectively. In addition, with a mixture containing both target bacteria in PBS and juice samples, the CE for S. aureus and B. cereus were recorded at about 90%, 91% and 80%, 84%, respectively. The MBs-OVA-Van plus mPCR method applied in PBS and juice samples for both bacteria showed limits of detection at 10 2 CFU mL -1 in a < 3 h total assay time. The results indicated that the method holds promise for isolation and enrichment of gram-positive bacteria with S. aureus and B. cereus as models.
研究领域:
微生物学
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生物技术
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分子生物学
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分析化学
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产品关联
DOI
高影响因子
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